The application of immune regenerative strategies to deal with unsolved pathologies, such as tendinopathies, is getting attention in the field of tissue engineering exploiting the innate immunomodulatory potential of stem cells [1]. In this context, Amniotic Epithelial Cells (AECs) represent an innovative immune regenerative strategy due to their teno-inductive and immunomodulatory properties [2], and because of their high paracrine activity, become a potential stem cell source for a cell-free treatment to overcome the limitations of traditional cell-based therapies. Nevertheless, these immunomodulatory mechanisms on AECs are still not fully known to date. In these studies, we explored standardized protocols [3] to better comprehend the different phenotypic behavior between epithelial AECs (eAECs) and mesenchymal AECs (mAECs), and to further produce an enhanced immunomodulatory AECs-derived secretome by exposing cells to different stimuli. Hence, in order to fulfill these aims, eAECs and mAECs at third passage were silenced for CIITA and Nrf2, respectively, to understand the role of these molecules in an inflammatory response. Furthermore, AECs at first passage were seeded under normal or GO-coated coverslips to study the effect of GO on AECs, and further exposed to LPS and/or IL17 priming to increase the anti-inflammatory paracrine activity. The obtained results demonstrated how CIITA and Nrf2 control the immune response of eAECs and mAECs, respectively, under standard or immune-activated conditions (LPS priming). Additionally, GO exposition led to a faster activation of the Epithelial-Mesenchymal transition (EMT) through the TGFβ/SMAD signaling pathway with a change in the anti-inflammatory properties. Finally, the combinatory inflammatory stimuli of LPS+IL17 enhanced the paracrine activity and immunomodulatory properties of AECs. Therefore, AECs-derived secretome has emerged as a potential treatment option for inflammatory disorders such as tendinopathies.
Digital Ventilated Cages (DVC) offer an innovative technology to obtain accurate movement data from a single mouse over time [1]. Thus, they could be used to determine the occurrence of a tendon damage event as well as inform on tissue regeneration [2,3]. Therefore, using the mouse model of tendon experimental damage, in this study it has been tested whether the recovery of tissue microarchitecture and of extracellular matrix (ECM) correlates with the motion data collected through this technology. Mice models were used to induce acute injury in Achilles tendons (ATs), while healthy ones were used as control. During the healing process, the mice were housed in DVC cages (Tecniplast) to monitor animal welfare and to study biomechanics assessing movement activity, an indicator of the recovery of tendon tissue functionality. After 28 days, the AT were harvested and assessed for their histological and immunohistochemical properties to obtain a total histological score (TSH) that was then correlated to the movement data. DVC cages showed the capacity to distinguish activity patterns in groups from the two different conditions. The data collected showed that the mice with access to the mouse wheel had a higher activity as compared to the blocked wheel group, which suggests that the extra movement during tendon healing improved motion ability. The histological results showed a clear difference between different analyzed groups. The bilateral free wheel group showed the best histological recovery, offering the highest TSH score, thus confirming the results of the DVC cages and the correlation between movement activity and structural recovery. Data obtained showed a correlation between TSH and the DVC cages, displaying structural and movement differences between the tested groups. This successful correlation allows the usage of DVC type cages as a non-invasive method to predict tissue regeneration and recovery.
Pressure ulcers are a common and recurrent clinical condition in paraplegic patients, requiring specialized equipment and care, as well as surgical interventions in order to treat them. This is especially true whenever and infection is declared, which will delay or impair ulcer epitelization. These surgical interventions require a good use of various myocutaneos flaps to cover all defects. Problem arises whenever there is not enough flap tissue to cover the entire ulcer, or when multiple surgeries to correct previous ulcers have already been performed. Our goal is to describe the use of a last resort surgical technique for covering infected pressure ulcers. This is a retrospective and descriptive case report based on data from clinical records, patient observation and analysis of complementary exams. We present the case of a 30-years-old man, paraplegic for 10 years due to motor vehicle accident with spinal cord injury. Since the accident, and although he used adapted equipment and pressure relief mattresses and wheelchair cushions, he developed recurrent, infected ulcers in the perineal and sacral area, being operated on for multiple times by the Plastic and Reconstructive Surgery (PRS) department, for surgical debridement and ulcer coverage with tensor faciae latae and hamstrings myocutaneous flaps. Despite all treatment, patient developed a stage IV perineal ulcer, which ranged from his left great trochanter to the right buttock, and a simultaneous stage IV sacral ulcer. Both ulcers were infected with meticilin-resistent Staphylococcus aureus (MRSA), sensitive to vancomycin. The patient's left hip joint was also infected (due to a direct trajectory to the perineal ulcer) and subluxated (due to absence of soft tissue support). A multidisciplinary team assembled and decision was made to disarticulate the patient's left hip, debride and irrigate extensively the surgical site, and use a double gastrocnemius myocutaneous fillet flap in-continuity, in a surgical collaboration between the Orthopaedics and PRS department. This should provide satisfactory soft tissue ulcer coverage as well as facilitate antibiotics penetrance and infection eradication. Surgery went without complications and the patient healed uneventfully. He resumed unrestricted positioning for sitting and wheelchair mobilization. Now, at two years follow-up, the patient still has no recurrence of either the ulcer or the infection. This surgical technique provided robust soft tissue coverage for the ulcers, as well as an important aid for infection control. It proved to be a viable option in a paraplegic patient, when more traditional flap techniques can no longer be used and with a recurrent infection.
Corynebacterium Jeikeium is a pathogen rarely involved in orthopaedic infections. Till date only 14 cases of osteomyelitis are described in the literature, envolving the tibia, foot and prosthethic (hip and joint) infection. To our knowledge, Corynebacterium Jeikeium as not been reported as an infectious agent of the spine. Our goal is to describe a case of scoliosis surgical site infection by a Corynebacterium Jeikeium specimen. This is a retrospective and descriptive case report based on data from clinical records, patient observation and analysis of complementary exams. We present a 24 year old female with a history of premature birth, West syndrome, spastic cerebral palsy and spina bifida. She was sent to our consult for evaluation of dorsolombar scoliosis. In October of 2014, she was submitted to surgery – posterior spine arthrodesis and instrumentation (D10 to L5) with bilateral pedicle screws and two chromium-cobalt bars. The early post-operative period was without complications. She was discharge at the seventh day of internment and was seen, fifteen days postoperative, at the consultation office, where the dressings were changed, with no signs of surgical site infection. One month post-operative, she recurs to the office because of an apparent seroma at the surgical site wound. There was no reference to fever or other signs of local/systemic infection. A swabbing of the wound was done and the patient was medicated with Ciprofloxacine, 500mg 12/12 hours – the culture came back negative. Seven days later she was seen again, maintaining the seroma with purge of a serous-aspect fluid. Antibiotic therapy was maintained and another swabbing was collected – culture came back negative. Because of suspected surgical site infection, she was re-operated at December of 2014. Surgical wound debridement was performed; three tissue samples and one exudate were sent to the microbiological department. In all samples but one was identified a Corynebacterium Jeikeium. No sensitivity test was performed. Intravenous Vancomicine, 1 gram 12/12 hours was started and maintained during 8 days. Eleven days post-operative she was discharged with oral Vibramicin, 100 milligrams 12/12 hours for two weeks. She is currently being followed at the doctor's office, with no sign of reinfection of the surgical site. This is the first reported case describing an infection of the spine by a Corynebactereium Jeikeium. Isolation of the causative agent of infection and literature-based directed antibiotherapy are important for a successful outcome.
Infection of the musculoskeletal (MSK) system is a dreaded complication that seems to be on the rise. Many factors, such as resistant bacteria or poor host factors, may influence such rise. This increase leads to greater resource consumption, especially due to antibiotic (ATB) prescription. Strategies must be created to detect patients at risk and prevent such infections. Hospital administrators should be made aware of the costs and impact of MSK infections in order to understand the need to prevent such complications. Therefore, our goal is to characterize the infected orthopaedic patient and evaluate the cost associated with ATB prescription in such cases. This is a retrospective and descriptive study, based on patient record analysis of all patients treated at our department, from January 2013 to March 2015. We identified 177 patients with a MSK infection and an isolated infectious agent. There was no predominance of either sex. Approximately 50% of patients were aged between 66 and 85 years old. Most frequent agents were MSSA (30,2%) and MRSA (21,8%), followed by Streptococcus species (8,9%) and Pseudomonas aeruginosas (7,1%). Although most patients were infected by a single agent, 18,6% had two or more causative agents isolated in cultures. Of those, 69,7% had either MSSA or MRSA as one of the isolated agents. Most commonly affected area was the hip (39,5%), followed by the knee (23,2%) and ankle (21,5%). Nearly all patients had had prior surgery, with arthroplasty and osteosynthesis as the most common procedures (52,4% and 33,3%, respectively). Infection related admission was responsible for a total of 6.116 hospital bed-days, with the majority of patients staying in hospital for up to 30 days (77,3%). Total cost of hospital ATB administration was 61.365,61€, with approximately 346,7€ spent in each patient. This corresponds to 42,6% of the total cost of medication during hospital stay (144.146,1€ of total cost, with 814,4€ per patient). The typical infected orthopaedic patient is aged 66 to 85 years old, was operated to his hip or knee, and either an arthroplasty or osteosynthesis was performed. Staphylococcus aureus remains the most common agent. MSK infections have significant costs associated with their treatment, and clinical departments should periodically re-evaluate discharged patients in order to understand what groups are at risk of developing such a complication. Only a correct identification of all these factors makes it possible to create a targeted preventive strategy, in order to reduce costs to the institution.
